AtPCaP1 | Arabidopsis thaliana plasma membrane cation-binding protein-1

AS10 931 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, R. sativus, B.rapa, B.oleracea

Product Information

Immunogen

KLH-conjugated peptide derived from Arabidopsis thaliana PCaP1 sequence UniProt: Q96262, TAIR: At4g20260

Host Rabbit

Clonality Polyclonal

Purity Serum

Format Lyophilized

Quantity 100 µl

Reconstitution For reconstitution add 100 µl of sterile water.

Storage Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

Tested applications Western blot (WB)

Recommended dilution 1 : 2000 (WB)

Expected | apparent MW

24.5 | 36 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana, Raphanus sativus (41 kDa), Brassica rapa (42 kDa), Brassica rapa var. glabra Regel (43 kDa), Brassica oleracea var. italica (41 kDa)

Predicted reactivity Brassicaceae
Species of your interest not listed? Contact us

Not reactive in Nicotiana benthamiana

Application examples

Additional information

Serum contains less than 0.1 % of sodium azide as preservative.

It is confirmed that the nock-out mutant (SALK_022955, Col-0 background) has no mRNA or protein of PCaP1. In the immunoblot, crude membrane fractions prepared from wild-type and pcap1 knock-out mutant plants were examined. Therefore the antibody to PCaP1 specifically recognizes PCaP1 and can be used for immunoblotting. For application in immunocytochemistry cross reacting band in proximity of Rubisco has to be removed by incubation of an antibody with a part of memrane with background band.

Protocol for preparation of crude membranes

This protocol will help to prepare crude membrane fractions from very small amount of plant tissue.
(1) Homogenize roots or root tips with the buffer without SDS.
(2) Centrifuge the homogenate in Eppendorf tubes at 2 000 rpm for 2 min to remove cell wall, nuclei, plastids and mitochondria.
(3) Centrifuge the obtained supernatant at 30 000 g for 20 min to separate soluble components. Please use adequate tubes for ultracentrifugation.
(4) Suspend the obtained precipitate (crude membranes) with SDS buffer with DTT or beta-mercaptoethanol.
(5) Heat the sample solution obtained the step (4) at 70 C for 5 min.
(6) Apply the denatured sample to SDS-PAGE.

Courtesy Prof. Msayoshi Maeshima, Nagoya University, Japan

Related products

antibodies to plasma membrane proteins

Plant protein extraction buffer

Secondary antibodies

Background

AtPCaP1 is a protein associated with plasma membrane with ability to bind Ca, calmodulin and phosphatidylinositol phoshphates. Alternative names: At4g20260, endomembrane-associated protein.

Product citations

Selected references Nagasaki et al. (2008) A hydrophilic cation-binding protein of Arabidopsis thaliana, AtPCaP1, is localized to plasma membrane via N-myristoylation and interacts with calmodulin and the posphatidylinositol phosphates PtdIns(3,4,5)P3 and PtdIns(3,5)P2. FEBS J. 275: 2267-2282.
Ide et al. (2007) Molecular properties of a novel, hydrophilic cation-binding protein associated with the plasma membrane. J. Exp. Botany, 58: 1173-1183.